Comparing the effects of chemical Ca2+ dyes and R-GECO on contractility and Ca2+ transients in adult and human iPSC cardiomyocytes
We compared commonly used BAPTA-derived chemical Ca<sup>2+</sup> dyes (fura2, Fluo-4, and Rhod-2) with a newer genetically encoded indicator (R-GECO) in single cell models of the heart. We assessed their performance and effects on cardiomyocyte contractility, determining fluorescent sign...
| Những tác giả chính: | , , , , , , , , , , , , , , , , |
|---|---|
| Định dạng: | Journal article |
| Ngôn ngữ: | English |
| Được phát hành: |
Elsevier
2023
|
| _version_ | 1826310940691267584 |
|---|---|
| author | Robinson, P Sparrow, AJ Psaras, Y Steeples, V Simon, JN Broyles, CN Chang, Y-F Brook, FA Wang, Y-J Blease, A Zhang, X Abassi, YA Geeves, MA Toepfer, CN Watkins, H Redwood, C Daniels, MJ |
| author_facet | Robinson, P Sparrow, AJ Psaras, Y Steeples, V Simon, JN Broyles, CN Chang, Y-F Brook, FA Wang, Y-J Blease, A Zhang, X Abassi, YA Geeves, MA Toepfer, CN Watkins, H Redwood, C Daniels, MJ |
| author_sort | Robinson, P |
| collection | OXFORD |
| description | We compared commonly used BAPTA-derived chemical Ca<sup>2+</sup> dyes (fura2, Fluo-4, and Rhod-2) with a newer genetically encoded indicator (R-GECO) in single cell models of the heart. We assessed their performance and effects on cardiomyocyte contractility, determining fluorescent signal-to-noise ratios and sarcomere shortening in primary ventricular myocytes from adult mouse and guinea pig, and in human iPSC-derived cardiomyocytes. Chemical Ca<sup>2+</sup> dyes displayed dose-dependent contractile impairment in all cell types, and we observed a negative correlation between contraction and fluorescence signal-to-noise ratio, particularly for fura2 and Fluo-4. R-GECO had no effect on sarcomere shortening. BAPTA-based dyes, but not R-GECO, inhibited in vitro acto-myosin ATPase activity. The presence of fura2 accentuated or diminished changes in contractility and Ca<sup>2+</sup> handling caused by small molecule modulators of contractility and intracellular ionic homeostasis (mavacamten, levosimendan, and flecainide), but this was not observed when using R-GECO in adult guinea pig left ventricular cardiomyocytes. Ca<sup>2+</sup> handling studies are necessary for cardiotoxicity assessments of small molecules intended for clinical use. Caution should be exercised when interpreting small molecule studies assessing contractile effects and Ca<sup>2+</sup> transients derived from BAPTA-like chemical Ca<sup>2+</sup> dyes in cellular assays, a common platform for cardiac toxicology testing and mechanistic investigation of cardiac disease physiology and treatment. |
| first_indexed | 2024-03-07T08:00:59Z |
| format | Journal article |
| id | oxford-uuid:c4249f10-f96b-43d8-bbdb-9cc25d3e02e8 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T08:00:59Z |
| publishDate | 2023 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | oxford-uuid:c4249f10-f96b-43d8-bbdb-9cc25d3e02e82023-09-26T11:26:41ZComparing the effects of chemical Ca2+ dyes and R-GECO on contractility and Ca2+ transients in adult and human iPSC cardiomyocytesJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:c4249f10-f96b-43d8-bbdb-9cc25d3e02e8EnglishSymplectic ElementsElsevier2023Robinson, PSparrow, AJPsaras, YSteeples, VSimon, JNBroyles, CNChang, Y-FBrook, FAWang, Y-JBlease, AZhang, XAbassi, YAGeeves, MAToepfer, CNWatkins, HRedwood, CDaniels, MJWe compared commonly used BAPTA-derived chemical Ca<sup>2+</sup> dyes (fura2, Fluo-4, and Rhod-2) with a newer genetically encoded indicator (R-GECO) in single cell models of the heart. We assessed their performance and effects on cardiomyocyte contractility, determining fluorescent signal-to-noise ratios and sarcomere shortening in primary ventricular myocytes from adult mouse and guinea pig, and in human iPSC-derived cardiomyocytes. Chemical Ca<sup>2+</sup> dyes displayed dose-dependent contractile impairment in all cell types, and we observed a negative correlation between contraction and fluorescence signal-to-noise ratio, particularly for fura2 and Fluo-4. R-GECO had no effect on sarcomere shortening. BAPTA-based dyes, but not R-GECO, inhibited in vitro acto-myosin ATPase activity. The presence of fura2 accentuated or diminished changes in contractility and Ca<sup>2+</sup> handling caused by small molecule modulators of contractility and intracellular ionic homeostasis (mavacamten, levosimendan, and flecainide), but this was not observed when using R-GECO in adult guinea pig left ventricular cardiomyocytes. Ca<sup>2+</sup> handling studies are necessary for cardiotoxicity assessments of small molecules intended for clinical use. Caution should be exercised when interpreting small molecule studies assessing contractile effects and Ca<sup>2+</sup> transients derived from BAPTA-like chemical Ca<sup>2+</sup> dyes in cellular assays, a common platform for cardiac toxicology testing and mechanistic investigation of cardiac disease physiology and treatment. |
| spellingShingle | Robinson, P Sparrow, AJ Psaras, Y Steeples, V Simon, JN Broyles, CN Chang, Y-F Brook, FA Wang, Y-J Blease, A Zhang, X Abassi, YA Geeves, MA Toepfer, CN Watkins, H Redwood, C Daniels, MJ Comparing the effects of chemical Ca2+ dyes and R-GECO on contractility and Ca2+ transients in adult and human iPSC cardiomyocytes |
| title | Comparing the effects of chemical Ca2+ dyes and R-GECO on contractility and Ca2+ transients in adult and human iPSC cardiomyocytes |
| title_full | Comparing the effects of chemical Ca2+ dyes and R-GECO on contractility and Ca2+ transients in adult and human iPSC cardiomyocytes |
| title_fullStr | Comparing the effects of chemical Ca2+ dyes and R-GECO on contractility and Ca2+ transients in adult and human iPSC cardiomyocytes |
| title_full_unstemmed | Comparing the effects of chemical Ca2+ dyes and R-GECO on contractility and Ca2+ transients in adult and human iPSC cardiomyocytes |
| title_short | Comparing the effects of chemical Ca2+ dyes and R-GECO on contractility and Ca2+ transients in adult and human iPSC cardiomyocytes |
| title_sort | comparing the effects of chemical ca2 dyes and r geco on contractility and ca2 transients in adult and human ipsc cardiomyocytes |
| work_keys_str_mv | AT robinsonp comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT sparrowaj comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT psarasy comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT steeplesv comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT simonjn comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT broylescn comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT changyf comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT brookfa comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT wangyj comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT bleasea comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT zhangx comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT abassiya comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT geevesma comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT toepfercn comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT watkinsh comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT redwoodc comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes AT danielsmj comparingtheeffectsofchemicalca2dyesandrgecooncontractilityandca2transientsinadultandhumanipsccardiomyocytes |